基于正交实验的赤苍藤组培快繁体系建立

doi:10.11983/CBB23040

摘要/Abstract

摘要： 以赤苍藤(Erythropalum scandens)带芽点的半木质化枝条为材料建立赤苍藤组培快繁体系, 研究外植体灭菌条件, 通过正交实验, 设计不同植物生长调节剂组合对赤苍藤愈伤组织诱导、愈伤组织分化、生根壮苗及移栽驯化的影响。结果表明, 赤苍藤外植体消毒最佳方法是以体积浓度为75%乙醇浸泡60秒后, 再以质量浓度为0.1% HgCl₂消毒10分钟, 成功率为48.89%; 叶片愈伤组织诱导的最佳培养基为MS+0.5 mg·L^-16-BA+1.0 mg·L^-12,4-D+1.0 mg·L^-1 IBA, 培养30天, 诱导率达71.11%, 且绿色紧密; 茎段诱导愈伤组织的最佳培养基为MS+1.0 mg·L^-16-BA+0.5 mg·L^-12,4-D+1.0 mg·L^-1IBA, 培养30天, 诱导率为70.00%, 且绿色紧密; 诱导愈伤组织增殖以及分化的最佳培养基为MS+2.0 mg·L^-16-BA+0.5 mg·L^-1TDZ+1.0 mg·L^-1 IBA, 芽分化率达98.89%, 增殖系数为3.33; 生根最佳培养基为MS+1.5 mg·L^-16-BA+0.5 mg·L^-1IBA, 生根率为100%, 平均生根数为2.2条; 生根苗在小颗粒泥炭土中成活率达88.89%。该研究建立了赤苍藤组培快繁体系, 为赤苍藤优质种苗生产奠定了良好基础。

关键词: 赤苍藤, 组培快繁, 无菌苗, 愈伤组织, 诱导与增殖

Abstract: To solve the problem of breeding excellent seedlings of Erythropalum scandens, research has been conducted on the establishment and optimization of tissue culture and rapid propagation systems of E. scandens by taking apical bud-induced aseptic seedlings as the material. Explant sterilization, callus induction, callus differentiation, test-tube rooting and transplanting and domestication were studied. The results are as follows: the best ratio of sterilization was 60 seconds of 75% alcohol+10 minutes of 0.1% HgCl₂, and the success rate was 48.89%. The best formula for callus induction by aseptic seedling leaf was MS+0.5 mg·L^-1 6-BA+1.0 mg·L^-1 2,4-D+1.0 mg·L^-1 IBA, for 30 days, and the induction rate was 71.11%, with compact green and strong differentiation potential. The best formula for callus induction by aseptic seedling shoot was MS+1.0 mg·L^-1 6-BA+0.5 mg·L^-1 2,4-D+1.0 mg·L^-1 IBA, for 30 days, and the induction rate was 70.00%; The most suitable medium for induction of callus propagation and differentiation was MS+2.0 mg·L^-16-BA+0.5 mg·L^-1 TDZ+1.0 mg·L^-1 IBA, bud differentiation rate was 98.89%, and coefficient of propagation was 3.33. The most suitable medium for rootage was MS+1.5 mg·L^-1 6-BA+0.5 mg·L^-1 IBA, achieving a 100% rootage rate with 2.2 of the average number. Plantlets were transplanted to small particle peat soil, and 88.89% of rooted plants survived. The research has established the tissue culture and rapid propagation system of E. scandens, which can be applied in production and serve as a foundation for providing seedlings and factory production.

Key words: Erythropalum scandens, tissue culture and rapid propagation, aseptic seedling, callus, induction and proli-feration

张尚文, 黄诗宇, 杨天为, 李婷, 张向军, 高曼熔. 基于正交实验的赤苍藤组培快繁体系建立. 植物学报, 2024, 59(1): 99-109.

Shangwen Zhang, Shiyu Huang, Tianwei Yang, Ting Li, Xiangjun Zhang, Manrong Gao. Establishment of a Tissue Culture and Rapid Propagation System for Erythropalum scandens Based on Orthogonal Test. Chinese Bulletin of Botany, 2024, 59(1): 99-109.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB23040

https://www.chinbullbotany.com/CN/Y2024/V59/I1/99

图/表 12

参考文献 37

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Treatment code	Contamination rate (%)	Survival rate (%)	Pollution causes
X-1	98.89±1.92 a	0.00±0.00 d	Fungal contamination
X-2	83.33±8.82 b	13.33±5.77 c	Fungal contamination
X-3	58.89±7.70 c	35.56±6.94 b	Fungal contamination
X-4	41.11±1.92 d	48.89±5.09 a	Fungal contamination
X-5	14.44±3.85 e	8.89±1.92 c	Fungal contamination

Treatment code	Contamination rate (%)	Survival rate (%)	Pollution causes
X-1	98.89±1.92 a	0.00±0.00 d	Fungal contamination
X-2	83.33±8.82 b	13.33±5.77 c	Fungal contamination
X-3	58.89±7.70 c	35.56±6.94 b	Fungal contamination
X-4	41.11±1.92 d	48.89±5.09 a	Fungal contamination
X-5	14.44±3.85 e	8.89±1.92 c	Fungal contamination

Plant growth regulator	Mass concentration (mg·L^-1)	Callus induction rate (%)		Callus condition
Plant growth regulator	Mass concentration (mg·L^-1)	Leaf	Shoot	Callus condition
IBA	0.1	13.33±5.77 b	18.89±1.92 b	Milky white, translucent
	0.2	33.33±6.67 a	22.22±5.09 b	Milky white, translucent
	0.4	28.89±3.85 a	38.89±3.85 a	Milky white, translucent
	0.6	28.89±1.92 a	36.67±5.77 a	Milky white, translucent
	0.8	20.00±3.33 b	35.56±8.39 a	Milky white, translucent
	1.0	5.56±1.92 d	4.44±1.92 cd	Milky white, translucent
	1.5	5.56±1.92 d	8.89±1.92 c	Milky white, translucent
	2.0	3.33±3.33 d	1.11±1.92 d	Milky white, translucent
	2.5	0.00±0.00 d	1.11±1.92 d	Stop growing
	3.0	0.00±0.00 d	0.00±0.00 d	Stop growing
2,4-D	0.1	20.00±5.77 c	17.78±1.92 d	Milky white, translucent
	0.2	22.22±6.94 c	22.22±3.85 cd	Milky white, translucent
	0.4	42.22±8.39 b	25.56±5.09 cd	Milky white, translucent
	0.6	47.78±3.85 b	28.89±3.85 c	Milky white, translucent
	0.8	42.22±3.85 b	38.89±9.62 b	Milky white, translucent
	1.0	48.89±8.39 b	45.56±5.09 b	Milky white, translucent
	1.5	27.78±1.92 c	38.89±1.92 b	Milky white, translucent
	2.0	75.56±6.94 a	28.89±3.85 c	Albinism, variation
	2.5	67.78±7.70 a	61.11±3.85 a	Albinism, variation
	3.0	1.11±1.92 d	5.56±9.62 e	Albinism, variation

Plant growth regulator	Mass concentration (mg·L^-1)	Callus induction rate (%)		Callus condition
Plant growth regulator	Mass concentration (mg·L^-1)	Leaf	Shoot	Callus condition
IBA	0.1	13.33±5.77 b	18.89±1.92 b	Milky white, translucent
	0.2	33.33±6.67 a	22.22±5.09 b	Milky white, translucent
	0.4	28.89±3.85 a	38.89±3.85 a	Milky white, translucent
	0.6	28.89±1.92 a	36.67±5.77 a	Milky white, translucent
	0.8	20.00±3.33 b	35.56±8.39 a	Milky white, translucent
	1.0	5.56±1.92 d	4.44±1.92 cd	Milky white, translucent
	1.5	5.56±1.92 d	8.89±1.92 c	Milky white, translucent
	2.0	3.33±3.33 d	1.11±1.92 d	Milky white, translucent
	2.5	0.00±0.00 d	1.11±1.92 d	Stop growing
	3.0	0.00±0.00 d	0.00±0.00 d	Stop growing
2,4-D	0.1	20.00±5.77 c	17.78±1.92 d	Milky white, translucent
	0.2	22.22±6.94 c	22.22±3.85 cd	Milky white, translucent
	0.4	42.22±8.39 b	25.56±5.09 cd	Milky white, translucent
	0.6	47.78±3.85 b	28.89±3.85 c	Milky white, translucent
	0.8	42.22±3.85 b	38.89±9.62 b	Milky white, translucent
	1.0	48.89±8.39 b	45.56±5.09 b	Milky white, translucent
	1.5	27.78±1.92 c	38.89±1.92 b	Milky white, translucent
	2.0	75.56±6.94 a	28.89±3.85 c	Albinism, variation
	2.5	67.78±7.70 a	61.11±3.85 a	Albinism, variation
	3.0	1.11±1.92 d	5.56±9.62 e	Albinism, variation

Mass concentration (mg·L^-1)	Callus differentiation rate (%)		Callus condition
Mass concentration (mg·L^-1)	6-BA	TDZ	6-BA	TDZ
0.1	4.44±1.92 ef	2.22±1.92 de	No change	Light green, 0-1 buds
0.2	6.67±3.33 e	10.00±5.77 e	No change	Light green, 0-1 buds
0.6	27.78±1.92 c	47.78±6.94 d	Light green	Light green, 0-2 buds
1.0	54.44±3.85 a	18.89±3.85 b	Light green, 1-2 buds	Light green, 0-3 buds
1.5	35.56±3.85 b	10.00±5.77 c	Light green, 1-2 buds	Light green,1-3 buds
2.0	23.33±5.77 cd	68.89±5.09 d	Light green, 2-3 buds	Partial albinism, 0-4 buds
2.5	18.89±3.85 d	68.89±1.92 a	Albinism, 3-6 buds	All albinism
3.0	3.33±3.33 ef	2.22±3.85 a	Albinism, 3-6 buds	Albinism
3.5	3.33±3.33 ef	0.00±0.00 e	Death	Death
4.0	0.00±0.00 f	0.00±0.00 e	Death	Death